Non-hydrogenated canola oil for food applications

ABSTRACT

A non-hydrogenated canola oil having superior oxidative stability and fry stability useful for food applications is disclosed, as well as seeds, plant lines and progeny thereof from which the oil is derived.

CROSS-REFERENCE TO RELATED APPLICATION

[0001] This application is a continuation-in-part application ofPCT/US94/04352, claiming priority under 35 USC §120, which is acontinuation-in-part application of Ser. No. 08/184,128 filed Jan. 21,1994, now abandoned, which is a continuation-in-part of Ser. No.08/054,806 filed Apr. 27, 1993, now abandoned.

FIELD OF THE INVENTION

[0002] The present invention relates to non-hydrogenated canola oilhaving improved flavor and performance attributes especially suitablefor food applications, and to the Brassica seeds, plant lines andprogeny thereof from which the oil is derived.

BACKGROUND

[0003] Canola oil has the lowest level of saturated fatty acids of allvegetable oils. As consumers become more aware of the health impact oflipid nutrition, consumption of canola oil in the U.S. has increased.However, generic canola oil has limited use in deep frying operations animportant segment of the food processing industry, due to itsinstability. Canola oil extracted from natural and commercial varietiesof rapeseed contains a relatively high (8%-10%) α-linolenic acid content(C_(18:3)) (ALA). The oil is unstable and easily oxidized duringcooking, which in turn creates off-flavors of the oil and compromisesthe sensory characteristics of foods cooked in such oils. It alsodevelops unacceptable off odors and rancid flavors during storage.

[0004] Hydrogenation can be used to improve performance attributes bylowering the amount of linoleic and α-linolenic acids in the oil. Inthis process the oil increases in saturated and trans fatty acids, bothundesirable when considering health implications. Blending of oil canalso be used to reduce the α-linolenic acid content and improve theperformance attributes. Blending canola oil with other vegetable oilssuch as cottonseed will increase the saturated fatty acids content ofthe oil but decreases the healthy attributes of canola oil.

[0005] α-Linolenic acid has been reported to oxidize faster than otherfatty acids. Linoleic and α-linolenic acids have been suggested asprecursors to undesirable odor and flavor development in foods. Toimprove the functionality of canola oil, the University of Manitobadeveloped the canola variety “Stellar” which has reduced α-linolenicacid (Scarth et al., Can. J. Plant Sci., 68:509-511 (1988)). The lowα-linolenic acid oil was reduced in odor when heated in air, but stillremained unacceptable to the sensory panel in flavor evaluations (Eskinet al., J. Am. Oil Chem. Soc. 66:1081-1084 (1989)). The oxidativestability of Stellar oil increased by 17.5% over the commercial varietyWestar as measured by Active Oxygen Method (AOM) hours. (Can. J. PlantSci. (1988.) Vol. 68, pp. 509-511).

[0006] European Patent Application, EP 0 323 753 A1 describes a canolaoil having an enhanced oleic acid content with increased heat stabilityin combination with other traits. The application further describes afrying oil with reduced α-linolenic acid which imparts increasedoxidative stability. No flavor and performance testing with thedescribed oil was reported.

[0007] Data which shows that oxidative stability is not solely relatedto fatty acid composition (described below) indicates that increasedstability cannot be inferred from fatty acid composition. The amount ofα-linolenic acid in the oil is only one factor which controls oxidativestability and flavor stability. Thus a canola oil which has improvedstability in its flavor and performance attributes for use in foodoperations is needed. The present invention provides such an oil.

SUMMARY OF THE INVENTION

[0008] The present invention provides an oil comprising anon-hydrogenated canola oil having an oxidative stability of from about37 to about 30 AOM hours in the absence of antioxidants. The oil of thepresent invention also has fry stability for up to at least 64 hours.After 64 hours of frying, the oil of the present invention has reducedtotal polar material content of about 23%, reduced free fatty acidcontent of about 0.7%, reduced red color development as shown by aLovibond color value of 6.7 red and reduced para-anisidine value of 125absorbance/g. After 32 hours of frying, the oil of the present inventionhas reduced total polar material content of about 12%, reduced freefatty acid content of about 0.3%, reduced red color development as shownby a Lovibond color of 2-0.7 red and reduced para-anisidine value of 112absorbance/g.

[0009] The present invention further provides a seed comprising aBrassica napus variety containing canola oil as described above, andprogeny thereof.

[0010] The present invention further provides a plant line comprising aBrassica napus canola variety which produces canola oil as describedabove, and individual plants thereof.

BRIEF DESCRIPTION OF THE SEED DEPOSIT

[0011] Seed designated IMC 130 as described hereinafter was depositedwith the American Type Culture Collection and was assigned accessionnumber 75446. Seed designated as A13.30137 as described hereinafter wasdeposited with the American Type Culture Collection and was assignedaccession number ______.

DETAILED DESCRIPTION OF THE INVENTION

[0012] The present invention provides canola oil having superior stableflavor and performance attributes when compared to known canola oils.The invention also provides Brassica napus seeds, and plant linesproducing seeds, from which such an oil can be produced.

[0013] A canola oil of the present invention is superior in oxidativestability and frystability compared to known canola oils. The superiorfunctionalities of the oil can be demonstrated, e.g., by standardizedAmerican Oil Chemists' Society (AOCS) oil testing methods. The improvedcharacteristics of the oil permit it to be used in new food products andpermit the oil to be used without hydrogenation in situations whereincreased flavor stability, oxidative stability, fry stability andshelf-life stability are desirable.

[0014] In the context of this disclosure, a number of terms are used. Asused herein, “functionality” or “performance attributes” meansproperties or characteristics of the canola oil and includes flavorstability, fry stability, oxidative stability, shelf-life stability, andphotooxidative stability.

[0015] Oxidative stability relates to how easily components of an oiloxidize which creates off-flavors in the oil, and is measured byinstrumental analysis using accelerated oxidation methods. American OilChemists' Society Official Method Cd 12-57 for Fat Stability: ActiveOxygen Method (re'vd 1989); Rancimat (Laubli, M. W. and Bruttel, P. A.,JOACS 63:792-795 (1986)); Joyner, N. T. and J. E. McIntyre, Oil and soap(1938) 15:184 (modification of the Schaal oven test). oils with highoxidative stability are considered to be premium oils for shelf stableapplications in foods, i.e., spray coating for breakfast cereals,cookies, crackers, fried foods such as french fries, and snack foodssuch as potato chips.

[0016] Fry stability relates to the resistance to degeneration of theoil during frying. Fry stability can be evaluated by measuringparameters such as total polar material content, free fatty acidcontent, color development and aldehyde generation. “Fry life” isdetermined by sequentially frying products in an oil and performing asensory analysis of the flavor of the fried products. Fry life ismeasured as the length of time the oil is used for frying before thesensory analysis of a fried product degrades to a predetermined score.Oils for restaurants, hospitals and large institutions primarily areused for frying foods and require fry stability.

[0017] Flavor stability is determined by sensory analysis of an oilsample periodically taken from an oil held under defined conditions. Forexample, oils may be stored in an oven at an elevated temperature toaccelerate the aging. The oil may also be stored at room temperature.However, the length of time required for testing renders this method tobe less useful. Flavor stability is measured by the time it takes forthe flavor of the oil to degrade to an established numerical score. Thesensory panel rates the oil or food product from 1 (unacceptable) to 9(bland). A rejection point is selected where the oil or food productbegins to show deterioration. Bottled cooking oils and salad dressingsrequire high flavor stability.

[0018] Photooxidative stability is determined from analysis of oilsamples taken periodically from oil stored under defined light andtemperature conditions. Photooxidative stability is reflected in theduration of time it takes for the flavor of the oil to degrade to a setscore. Bottled cooking oils require high photooxidative stability.

[0019] Shelf-life stability is determined by the analysis of foodsamples cooked in the oil, then packaged and stored in an oven at anelevated temperature to accelerate aging. “Shelf-life” is the time ittakes for the flavor of the food to degrade to give a set score oils forfried snacks require shelf-life stability.

[0020] As used herein, a “line” is a group of plants that display littleor no genetic variation between individuals for at least one trait ofinterest. Such lines may be created by several generations ofself-pollination and selection, or vegetative propagation from a singleparent using tissue or cell culture techniques. As used herein, theterms “cultivar” and “variety” are synonymous and refer to a line whichis used for commercial production.

[0021] “Saturated fatty acid” refers to the combined content of palmiticacid and stearic acid. “Polyunsaturated fatty acid” refers to thecombined content of linoleic and α-linolenic acids. The term “room odor”refers to the characteristic odor of heated oil as determined using theroom-odor evaluation method described in Mounts (J. Am. Oil Chem. Soc.,56:659-663, 1979).

[0022] A “population” is any group of individuals that share a commongene pool. The term “progeny” as used herein means the plants and seedsof all subsequent generations resulting from a particular designatedgeneration.

[0023] The term “selfed” as used herein means self pollinated.

[0024] “Generic canola oil” refers to a composite blend of oilsextracted from commercial varieties of rapeseed currently known, whichvarieties generally exhibited at a minimum 8-10% α-linolenic acidcontent, a maximum of 2% erucic acid and a maximum of 30 μmol/g totalglucosinolate level. The seed from each growing region is graded andblended at the grain elevators to produce a uniform product. The blendedseed is then crushed and refined, the resulting oil being a blend ofvarieties and sold for use. Table 1 shows the distribution of canolavarieties seeded as percentage of all canola seeded in Western Canada in1990. Canada is a leading producer and supplier of canola seed and oil.TABLE 1 Distribution of Canola Varieties Grown in Western Canada in 1990Canola Variety Percent of Seeded Area A. campestris Candle 0.4 Colt 4.4Horizon 8.5 Parkland 2.5 Tobin 27.1 B. napus Alto 1.1 Delta 0.9 Global0.9 Legend 18.2 Pivot 0.1 Regent 0.5 Stellar 0.2 Tribute 0.4 Triton 0.7Triumph 0.2 Westar 29.5 Others 4.4

[0025] “Canola” refers to rapeseed (Brassica) which has an erucic acid(C₂₂₁) content of at most 2 percent by weight based on the total fattyacid content of a seed, preferably at most 0.5 percent by weight andmost preferably essentially 0 percent by-weight and which produces,after crushing, an air-dried meal containing less than 30 micromoles(μmol) per gram of defatted (oil-free) meal.

[0026] The term “canola oil” is used herein to describe an oil derivedfrom the seed of the genus Brassica with less than 2% of all fatty acidsas erucic acid.

[0027] Genetic crosses are made with defined germplasm to produce thecanola oil of the present invention having reduced polyunsaturated fattyacids, improved flavor stability, fry stability, oxidative stability,photooxidative stability and shelf-life stability, in a high yieldingspring canola background. IMC 129, a Spring canola variety with higholeic acid in the seed oil is crossed with IMC 01, a spring canolavariety with low α-linolenic acid in the seed oil. Flower buds of the F₁hybrid are collected for microspore culture to produce a dihaploidpopulation. The dihaploid plants (genetically homozygous) are selectedwith high oleic, and reduced linoleic and α-linolenic acids in the seedoil and field tested for stability of the fatty acids and yield.

[0028] After five generations of testing in the field and greenhouse ahigh yielding selection with fatty acid stability in multipleenvironments is selected. Seed of selection is grown in isolation,harvested, and the oil extracted and processed to produce a refined,bleached and deodorized oil using known techniques. The oil produced wasfound to be functionally superior in oxidative stability and frystability relative to a commercial-type, generic canola oil processedunder similar conditions.

[0029] The canola oil of the present invention has an oxidativestability as determined by Accelerated Oxygen Method (AOM) values offrom about 35 to about 40 hours. This is significantly higher than anyknown pilot plant or commercial processed canola oil. The increase is 45to 60% above commercial type generic canola oil.

[0030] Under extended frying conditions, canola oil of the invention issignificantly lower than commercial-type generic canola in the oxidativetests for total polar material, free fatty acids, color development andp-anisidine value. The oil remains significantly lower in all oxidativeparameters tested after 32 and 64 hours of frying.

[0031] The oil has about 12% and about 23% total polar materials at 32and 64 hours of frying, respectively. This represents a 34% decrease at32 hours and a 17% decrease at 64 hours compared to commercial typegeneric canola oil. The total polar materials are a measure of the totalamount of secondary by-products generated from the triacylglycerols as aconsequence of oxidations and hydrolysis, and their reduction indicatesimproved oxidative stability.

[0032] Oil of the invention has a reduced content of free fatty acids ofabout 0.3% and about 0.7% at 32 and 64 hours of frying, respectively.This represents a 37% decrease at 32 hours and a 23% decrease at 64hours compared to commercial type generic canola oil. The level of freefatty acids is a measure of oxidation and hydrolysis of thetriacylglycerols and their reduction also indicates improved oxidativestability.

[0033] The color developed in an oil during frying is also an indicationof triacylglycerol oxidation. The oil of the present inventiondemonstrated a reduced level of color development. The Lovibond color isabout 2.7 red and about 6.7 red at 32 and 64 hours of frying,respectively. This represents a 38% decrease at 32 hours and a 47%decrease at 64 hours compared to commercial type generic canola oil.

[0034] Reduced development of aldehydes during frying also indicateimproved oxidative stability and are measured by the p-anisidine valuein absorbance/g at 350 nm. The oil has a p-anisidine value of about 112absorbance/g after 0.32 hours of frying and of about 125 absorbance/gafter 64 hours of frying. This represents a 32% decrease at 32 hours anda 14% decrease at 64 hours compared to commercial type generic canolaoil.

[0035] The oil additionally has improved oxidative stability and fryingstability without hydrogenation or the addition of antioxidants. Theimproved oxidative and fry stability results in increased flavorstability of the oil. Addition of antioxidants to the oil will furtherincrease oxidative stability.

[0036] Oil of the invention may be produced from, for example, aBrassica napus plant designated as IMC 130 or from a Brassica napus linedesignated as A13.30137. The seed oil has reduced amounts of totalC_(16:0) (palmitic) and C_(18:0) (stearic) saturates of less than 6.5%,oleic acid from 74 to 80%, linoleic acid from 5 to 12%, α-linolenic acidfrom 2.0 to 5.0% and erucic acid of less than 1%.

[0037] The oil of the present invention is especially suitable for usein food applications, in particular for frying foods, due to itssuperior oxidative stability and fry stability. Due to itsnon-hydrogenated nature, it is especially desirable for positive humanhealth implications. The seeds, plant lines, and plants of the presentinvention are useful for the production of the non-hydrogenated canolaof this invention.

EXAMPLES

[0038] The present invention is further defined in the followingExamples, in which all parts and percentages are by weight and degreesare Celsius, unless otherwise stated. It should be understood that theseExamples, while indicating preferred embodiments of the invention, aregiven by way of illustration only.

Example 1

[0039] A cross of IMC 129×IMC 01 was conducted to obtain A13.30038, adihaploid Spring canola variety. IMC 129 (U.S. PVP Certificate No.9100151) is a Spring canola Brassica napus variety possessing high oleicacid (>75%) in the seed oil. IMC 01 is a Spring canola Brassica napusvariety possessing low α-linolenic acid (<2.5%) in the seed oil. Agenetic cross was made in 1989 to combine the low α-linolenic and higholeic acid traits in a high yielding background for commercialproduction.

[0040] The. F₁ plants (IMC 129×IMC 01) were grown in a growth chamber at12°/6° C. (day/night) with 16 hours of illumination. Flower buds between2-3.5 mm were selected for microspore isolation. The microspores wereisolated and cultured to produce embryos using the method of Lichter,R., Z. Pflanzenphysiol, 105:427-434 (1982). Plants regenerated from themicrospores were grown in the greenhouse until flowering. Haploid plantswere treated with colchicine to induce chromosome doubling. Dihaploidplants were self-pollinated.

[0041] Seed (DH₁) of the selfed dihaploid plants were harvested in 1990and analyzed in bulk for fatty acid composition via gas chromatography.Following fatty acid analysis, seed designated A13.30038 was identifiedwith high oleic and low α-linolenic acids (Table 2). The DH₁ seed wasplanted in the greenhouse and self-pollinated. The harvested DH₂ seedgeneration maintained the selected fatty acid composition. In 1991 theDH₂ seed was planted in Southeast Idaho to determine yield of the line.Plants in the field were self-pollinated to determine fatty acidstability. The DH seed maintained the selected fatty acid composition inthe field. DH₃ seed of A13.30038 was increased under isolation tentsduring the Winter of 1991 in Southern California. The DH₄ seedmaintained the selected fatty acid stability and was increased inisolation (2 miles) in Southeastern Idaho during 1992 to further testoil quality and yield.

[0042] After the 1992 summer trial, A13.30038 was found to have improvedyield and stable fatty acid composition. This line was renamed IMC 130.Yield data for line IMC130 is shown in Tables 13 and 14; the fatty acidcomposition over five generations is listed in Table 2. TABLE 2 FattyAcid Composition of A13.30038 over Five Generations Fatty AcidComposition Generation C_(16:0) C_(18:0) C_(18:1) C_(18:2) C_(18:3)C_(22:1) DH₁ 3.8 2.8 80.5 6.6 2.3 0.0 DH₂ 3.8 2.8 77.8 8.4 3.3 0.0 DH₃3.6 2.2 80.0 7.5 2.8 0.0 DH₄ 3.3 1.9 79.3 8.9 3.6 0.0 DH₅ 3.6 2.9 76.210.3 3.4 0.0

[0043] The IMC 130 seed was crushed and the resulting oil processedusing a pilot plant at the POS Pilot Plant Corporation, 118 VeterinaryRoad, Saskatoon, Saskatchewan, Canada. The starting seed weight was 700kg. All oils for which data is supplied were prepared under the sameconditions set forth below. To ensure good extraction of the oil theseed was tempered by spraying the seed with water to raise the moistureto 8.5%. The seed and water were blended and allowed to equilibrate. Theseed was flaked using smooth roller. A minimum gap setting was used toproduce a flake thickness of 0.23 to 0.27 mm. The oil cells were furtherruptured and enzymes deactivated by heating in a two tray cooker. Thetop tray was heated to 65-75° C. and the bottom tray to 91-95° C. usingdry heat. A sweeping arm was used to agitate the material.

[0044] The oil was pressed from the flaked seed using a Simon-Rosedowns9.5 cm diameter by 94 cm long screw press operating at a screw speed of17 rpm. The crude oil was kept under nitrogen until further processing.Hexane extraction was used to remove the oil from the press cake. Thepress cake was extracted using a total residence time of 90 min. and asolvent to solids ration of 2.4:1. The crude solvent extracted oil wascollected and kept under nitrogen until further processing.

[0045] The crude press and solvent extracted oils were dried andfiltered to remove solids prior to degumming. The oils were heated at100° C. under a vacuum until foaming stopped. The oil was cooled to65-75° C. and 0.8% of filter aid was added and filtered. The oil waswater degummed to remove phosphatides from the oil. The blended pressand solvent extracted oils were heated to 60-70° C. and 2.0% of 80-100°C. water was added and mixed for 15 min. The oil was then centrifugedfor gum removal.

[0046] Alkali refining was used to remove the free fatty acids.Phosphoric acid (85%) was added to 0.25% of the water degummed oil heldat 60-70° C. and mixed for 30 min. sodium hydroxide was added to theacid treated oil to neutralize the free fatty acids. After a 15 min.retention time the oil was heated to 75-80° C. and centrifuged.

[0047] Water washing was done to further remove soaps. The refined oilwas washed by adding 15% of 90-95° C. water to the oil and mixed for 15min. The oil was maintained at 75-80° C. and centrifuged. The washed oilwas heated to 60-65° C. and bleached using Englehard's ‘Grade 160’bleaching clay. The oil was heated to 105-110° C. and held under avacuum for 30 min. The oil was cooled to 60-65° C. and 20% of the clayweight was added as a filter aid.

[0048] The bleached oil was deodorized using a Johnson-Loft packed towercontinuous deodorizer. The oil deodorization temperature was 265° C. andthe feed rate was 200 kg/hr. The steam rate was 1% of the feed rate andthe system pressure was 0.16-0.18 kPa. The oil was preheated to 68-72°C. prior to being fed into the deareation vessel. The oil was cooled to41-42° C. prior to removal of the vacuum. The oil was stored undernitrogen at −20° C.

[0049] The IMC 130 oil was analyzed for fatty acid composition via gaschromatography along with commercially available canola oils. Table 3provides data on the fatty acid profiles of IMC 130 oil compared tocommercially available canola oils, IMC 129 (a high oleic acid oil), IMC144 (a typical generic canola oil) and Brand A (a typical generic canolaoil). The data demonstrates reduced levels of linoleic (C_(18:2)),α-linolenic (C_(18:3)), and total polyunsaturated fatty acids for IMC130. TABLE 3 Fatty Acid Composition of Refined, Bleached and DeodorizedOils Fatty Acid Composition (%) Variety C_(16:0) C_(18:0) C_(18:1)C_(18:2) C_(18:3) Total Polys* IMC 130 3.6 2.9 76.2 10.3 3.4 13.7 IMC144 2.9 2.1 62.6 19.5 8.1 27.6 IMC 129 3.9 2.0 78.8 7.7 3.9 11.6 Brand A3.8 2.0 60.9 19.9 9.1 28.0

[0050] Oils were evaluated for AOM hours under the methods outlined inthe American Oil Chemists' Society (AOCS) Official Method Cd 12-57 forFat Stability: Active Oxygen Method (re'vd. 1989). The degree ofoxidative stability is rated as the number of hours to reach a peroxidevalue of 100. Each oil sample was prepared in duplicate.

[0051] The IMC 130 oil was found to have significantly higher AOM hoursthan other oils tested, IMC 144, IMC 01, and IMC 129 after similar pilotplant processing. The IMC 144, IMC 0.01, and IMC 129 oils are currentlycommercially available from InterMountain Canola, Cinnaminson, N.J.(Table 4). IMC 129 (a high oleic variety) and IMC 01 (a low α-linolenicvariety) were the parent lines crossed to generate IMC 130. IMC 144 is atypical generic canola oil. IMC 130 oil has a minimum of 37 AOM hourswhich is significantly greater than commercial-type, generic canola at22 AOM hours. Typically, pilot plant processing of oils tends to reduceAOM hours as the process is much harsher on the oil than commercialprocessing. The greater oxidative stability of IMC 130 can be attributedto a lower polyunsaturated fatty acid content than the IMC 144 oil ontypical generic canola oil (Table 3). The greater oxidative stabilityover the high oleic IMC 129 oil, which is similar in fatty acidcomposition to IMC 130 oil, indicates that oxidative stability is notsolely related to fatty acid content. TABLE 4 AOM Hours of Pilot PlantProcessed Canola Oils Process Method IMC 144 IMC 01 IMC 129 IMC 130(Generic) (Low ALA)* (High Oleic) (Example 1) Pilot Plant 15-22 20-22 1637-40

Example 2

[0052] The oil of Example 1 and IMC 144, a generic canola oil, weresubjected to further testing to determine frying stability as measuredby oxidative degradation during frying.

[0053] 1900 g of each test oil was placed in a clean six quart capacity110 volt, commercial fryer (Tefal Super Cool Safety Fryers Model 3617).Oil temperature was maintained at 190° C. for eight hours each day.Temperature was controlled to +5° C. of the target temperature using aCole-Palmer temperature controller.

[0054] Commercially available frozen french fries (100 g) were fried forfour min, three times per eight hour day in each test oil. 50 mL of oilwere removed each day for chemical analysis to determine the amount ofoxidative degradation. Fresh oil was added to the fryer each day toreplace the amount removed for samples or lost: through absorption onfries and retention on process equipment.

[0055] The oxidative parameters of the oils after frying were measuredusing procedures established by the AOCS (Official Methods andRecommended Practices of the American Oil Chemists' Society, FourthEdition (1989) Ed., D. Firestone, Published by the American OilChemists' Society, Champaign, Ill.). These oxidative parameters areindicators of fry stability.

[0056] The oil was analyzed after frying for Total Polar Material(%TPM), Free Fatty Acids (%FFA), Color Development and Para-AnisidineValue (P-AV). The resulting data at 0, 32, and 64 hours of frying isreported in Table 5. The IMC 130 values reported in Table 5 aresignificantly lower than the IMC 144 values with a 95% degree ofconfidence. This demonstrates the improved fry stability of IMC 130 oilover commercial IMC 144 canola oil.

[0057] The percent of total polar material was determined using the AOCSOfficial Method Cd 20-91. The total polar materials are a measure of thetotal amount of secondary by-products generated from thetriacylglycerols as a consequence of oxidations and hydrolysis. Reducedaccumulation of total polar material by an oil indicates improvedoxidative stability. IMC 130 was significantly reduced in total polarmaterial after 32 and 64 hours of frying in comparison to commercialcanola oil.

[0058] The percent of free fatty acids were determined using AOCSOfficial Method Ca 5a0-40. Free fatty acids generated in the oil duringfrying is a measure of oxidation and hydrolysis of the triacylglycerols.Reduced free fatty acids during frying of an oil indicates improvedoxidative stability. In comparison to commercial canola oil, IMC 130 oilwas significantly reduced in free fatty acids after 32 and 64 hours offrying.

[0059] Color development was measured using the AOCS Official Method Cc13b-45 using a Lovibond Tintometer and is reported as red color. Redcolor development in the oil during frying is an indication oftriacylglycerol oxidation. Oils with reduced red color development willhave improved oxidative stability. IMC 130 oil had significantly lessred color development than the commercial oil after 32 and 64 hours offrying.

[0060] The para-anisidine value was measured using the AOCS OfficialMethod Cd 18-90. Aldehydes are generated during frying from theoxidation of the triacylglycerol are measured by the p-anisidine value.The p-anisidine value is 100 times the optical density measured at 350nm in a 1 cm cell of a solution containing 1.00 g of the oil in 100 μLof a mixture of solvent and reagent according to the method referenced,and is in absorbance/g. Reduced development of aldehydes during fryingis an indicator of improved oxidative stability of the oil. IMC 130 hadsignificantly less aldehyde content after 32 and 64 hours of frying thanthe IMC 144 canola oil, a typical commercial generic canola oil. TABLE 5Effects of Frying on Oxidative Parameters IMC IMC IMC IMC IMC IMCOxidation 130 at 144 at 130 at 144 at 130 at 144 at Parameter 0 Hrs. 0Hrs. 32 Hrs. 32 Hrs. 64 Hrs. 64 Hrs. % TPM^(a) 5.3 5.8 12.0 18.2 22.627.2 % FFA^(b) 0.01 0.01 0.29 0.46 0.74 0.96 Lovibond 0.3 0.5 2.7 4.36.7 12.5 Color^(c) P-AV^(d) 0.27 1.65 112 164 125 145

Example 3

[0061] The oil of Example 5 plus the following oils were subjected tofurther testing.

[0062] IMC 129—high oleic canola oil

[0063] Quality analysis of each oil is found in Table 6. TABLE 6 OilAnalysis IMC 130 IMC 129 Red Color 0.8 0.3 Yellow Color 6 2para-anisidine value³ 2.58 0.66 Peroxide Value¹ 0.3 0.3 Totox Value²3.18 1.24 % Polars 0.69 .64 % Polymers 0.013 0.010 % Free Fatty Acids0.022 0.014 % C16:0 3.5 3.6 % C18:0 2.3 2.0 % C18:1 73.4 75.7 % C18:211.1 9.5 % C18:3 5.7 6.2

[0064] Oxidative stability of the oil in Example 5 was demonstrated bymeasuring the increase in Peroxide Value and in para-Anisidine Valuegenerated under accelerated aging conditions using a modified Schaaloven test. The test oil (200 g) was placed in an 500 ml uncovered amberglass bottle with a 4.3 cm opening, and placed in a 60° C. convectionoven. One bottle was prepared for each evaluation. Results are found inTable 7 and Table 8.

[0065] The peroxide value was measured using the AOCS Official Method Cd8b-90. Hydroperoxides generated from oxidation of the triacylglycerolswere measured by the peroxide value. The peroxide value was expressed interms of milliequivalents of peroxide per 1000 grams of sample (meq/Kg).Reduced development of hydroperoxides during storage was an indicator ofimproved oxidative stability.

[0066] The para-anisidine value was measured using the AOCS OfficialMethod Cd 18-90. Aldehydes generated from the oxidation of thetriacylglycerol was measured by the p-anisidine value. The p-anisidinevalue was 100 times the optical density measured at 350 nm in a 1 cmcell of a solution containing 1.00 g of the oil in 100 ml of a mixtureof solvent and reagent according to the method referenced, and isabsorbance/g. Reduced development of aldehydes during storage was anindicator of improved oxidative stability of the oil. TABLE 7Accelerated Aging - Oxidative Stability Increase in Peroxide Value,Milliequivalents per kg Days in oven: IMC 130 IMC 129 3 0.9 0.7 6 2.12.3 9 12.6 14.9 12 16.1 22.1 15 24.5 29.7

[0067] TABLE 8 Accelerated Aging - Oxidative Stability Increase inpara-Anisidine Value, Absorbance per g Days in oven: IMC 130 IMC 129 60.1 0.2 9 2.0 3.1 12 4.8 6.9 15 6.9 10.2

Example 4

[0068] The oil of Example 5 plus the following oils were tested foroxidative stability.

[0069] Brand T—commercially available high oleic sunflower oil

[0070] Brand A—commercially available generic canola oil

[0071] Quality analysis of each oil is found in Table 9.

[0072] Oxidative stability of the oils described in Table 9 weredetermined under accelerated aging conditions using a modified Schaaloven test which accelerates oxidation. Oxidative stability wasdemonstrated by the increase in peroxide value over the test period.Schaal oven tests show that each day of accelerated oxidation at 60° C.is equivalent to a month of oxidation under ambient storage conditions.Using this correlation three days of accelerated aging is equivalent tothree months of ambient storage. The test oil (200 g) was placed in a500 ml uncovered amber glass bottle with a 4.3 cm opening, and placed ina 60° C. convection oven. One bottle was prepared for each evaluation.The test was carried out to six days to simulate actual productshelf-life of six months. Results are found in Table 10.

[0073] The peroxide value was measured using the AOCS Official Method Cd8b-90. Hydroperoxides generated from oxidation of the triacylglycerolswere measured by the peroxide value. The peroxide value was expressed interms of milliequivalents of peroxide per 1000 grams of sample (meq/Kg).Reduced development of hydroperoxides during storage is an indicator ofimproved oxidative stability. IMC 130 had significantly less peroxidedevelopment after three days and six days in the Schaal oven test thanBrand T, a high oleic sunflower oil with lower polyunsaturates(C_(18:2)+C_(18:3)), and Brand A, a typical commercial generic canolaoil. TABLE 9 Quality Analysis of Test Oils IMC 130 Brand T Brand A RedColor 0.8 0.8 0.7 Yellow Color 6 6 5 para-anisidime value³ 2.58 4.212.32 Peroxide Value¹ 0.3 0.6 0.7 Totox Value² 3.18 5.41 3.72 % Polars0.69 0.90 0.36 % Polymers 0.013 0.004 0.01 % Free Fatty Acids 0.0220.016 0.013 % C16:0 3.5 3.3 4.0 % C18:0 2.3 4.2 2.0 % C18:1 73.4 81.762.5 % C18:2 11.1 8.8 18.3 % C18:3 5.7 0.3 7.7

[0074] TABLE 10 Accelerated Aging - Oxidative Stability Increase inPeroxide Value, Meq/Kg Days in Oven: IMC 130 Brand A Brand T 3 0.8 6.23.2 6 1.8 14.4 7.0

Example 5

[0075] IMC 130 canola seed was produced during the 1993 growing seasonof the Northwestern U.S. The resulting IMC 130 canola seed was cleanedthrough commercial seed cleaning equipment to remove foreign matterconsisting of weed seeds, canola plant material immature canola seed andother non canola matter.

[0076] The cleaned IMC 130 canola seed was crushed and the resulting oilwas processed at SVO Specialty Products, Inc., One Mile East,Culbertson, Mont. Approximately 361 tons of IMC 130 canola seed wascrushed under the processing conditions outlined below.

[0077] Whole canola seed was passed through a double roll Bauermeisterflaking rolls with smooth surface rolls available from BauermeisterInc., Memphis, Tenn. 38118. The roll gap was adjusted so as to produce acanola flake 0.25 to 0.30 mm thickness. Flaked canola seed was conveyedto a five tray, 8 foot diameter stacked cooker, manufactured by CrownIron Works, Minneapolis, Minn. 55440. The flaked seed moisture wasadjusted in the stacked cooker to 5.5-6.0%. Indirect heat from the steamheated cooker trays was used to progressively increase the seed flaketemperature to 80-90° C., with a retention time of approximately 20-30minutes. A mechanical sweep arm in the stacked cooker was used to ensureuniform heating of the seed flakes. Heat was applied to the flakes todeactivate enzymes, facilitate further cell rupturing, coalesce the oildroplets and agglomerate protein particles in order to ease theextraction process.

[0078] Heated canola flakes were conveyed to a screw press from AndersonInternational Corp., Cleveland, Ohio 44105 equipped with a suitablescrew worm assembly to reduce press out approximately 70% of the oilfrom the IMC 130 canola flakes. The resulting press cake contained15.0-19.0% residual oil.

[0079] Crude oil produced from the pressing operation was passed througha settling tank with a slotted wire drainage top to remove the solidsexpressed out with the oil in the screw pressing operation. Theclarified oil was passed through a plate and frame filter to remove theremaining fine solid canola particles. The filtered oil was combinedwith the oil recovered from the extraction process before oil refining.

[0080] Canola press cake produced from the screw pressing operation wastransferred to a FOMM basket extractor available from French Oil Milland Machinery Co., Piqua, Ohio 45356, where the oil remaining in thecake was extracted with commercial n-Hexane at 55° C. Multiple countercurrent hexane washes were used to substantially remove the remainingoil in the press cake, resulting in a press cake which contained1.2-2.3%, by weight, residual oil in the extracted cake. The oil andhexane mixture (miscella) from the extraction process was passed througha two stage rising film tube type distillation column to distill thehexane from the oil. Final hexane removal from the oil was achieved bypassing the oil through a stripper column containing disk and doughnutinternals under 23-26 in. Hg vacuum and at 107-115° C. A small amount ofstripping steam was used to facilitate the hexane removal. The canolaoil recovered from the extraction process was combined with the filteredoil from the screw pressing operation, resulting in blended crude oil,and was transferred to oil processing.

[0081] In the oil processing the crude oil was heated to 66° C. in abatch refining tank to which 0.15% food grade phosphoric acid, as 85%phosphoric acid, was added. The acid serves to convert the nonhydratable phosphatides to a hydratable form, and the chelate minormetals that are present in the crude oil. The phosphatides and the metalsalts are removed from the oil along with the soapstock. After mixingfor 60 minutes at 66° C., the oil acid mixture was treated withsufficient sodium hydroxide solution (12° Be) to neutralize the freefatty acids and the phosphoric acid in the acid oil mixture. Thismixture was heated to 71° C. and mixed for 35 minutes. The agitation wasstopped and the neutralized free fatty acids, phosphatides, etc.(soapstock) were allowed to settle into the cone bottom of the refiningtank for 6 hours. After the settling period, the soapstock was drainedoff from the neutralized oil.

[0082] A water wash was done to reduce the soap content of the oil byheating the oil to 82° C. and by adding 12% hot water. Agitation of themixture continued for 10 minutes. The mixture was allowed to settle outfor 4 hours at which time the water was drained off the bottom of therefining vessel.

[0083] The water washed oil was heated to 104-110° C. in a vacuumbleacher vessel maintained at 24-26 in. Hg vacuum. A slurry of the IMC130 canola oil and Clarion 470 bleaching clay available from AmericanColloid Company, Refining Chemicals Division, Arlington Heights, Ill.60004, was added to the oil in the vacuum bleacher. This mixture wasagitated for 20 minutes before filtering to remove the bleaching clay.The clay slurry addition was adjusted to provide a Lovibond color AOCSOfficial Method Cc 136-4 of less than 1.0 red units when the oil washeated to 288° C. under atmospheric pressure. Nitrogen was injected intothe filtered bleached oil and maintained under a nitrogen blanket untilthe oil was deodorized.

[0084] Refined and bleached IMC 130 canola oil was deodorized in asemi-continuous Votator deodorizer tower at a rate of approximately7,000 pounds per hour. The deodorization temperature was maintained at265-268° C. with a system pressure of 0.3-0.5 mm Hg absolute pressure.Approximately 1-1.5% sparge steam was used to strip off the free fattyacids, color bodies, and odor components. Retention time in thedeodorizer was 50-70 minutes. The deodorized oil was cooled to 45-50° C.and nitrogen was injected prior to removal of the vacuum. The deodorizedoil was stored under a nitrogen blanket.

[0085] The resulting IMC 130 deodorized oil was analyzed for fatty acidcomposition via gas chromatography. The percent fatty acids wereC_(16:0) of 3.6%, C_(18:0) of 2.2%, C_(18:1) of 74.3% C_(18:2) of 11-9%,C_(18:3) of 4.8% and total polyunsaturated of 16.7%. These data can becompared to the values for IMC 144, IMC 129 and Brand A as shown inTable 3. The data demonstrates that IMC 130 maintains reduced levels oflinolenic acid (C_(18:2)), α-linolenic (C_(18:3)), and totalpolyunsaturated fatty acids when compared to typical generic canola oilsIMC 144 and Brand A.

[0086] Table 11 provides data on the AOM hours of the IMC 130 oilprocessed as described above (the commercial process in 1993), comparedto commercially available canola oils, IMC 129 (a high oleic acid oil),IMC 144 (a typical generic canola oil) and IMC 01 (a low α-linolenicoil). The IMC 130 oil was evaluated for AOM hours under the methodsoutlined in the American Oil Chemists' (AOCS) Official Method Cd 12-57for Fat Stability: Active Oxygen Method (re'vd 1989). The degree ofoxidative stability is rated as the number of hours to reach a peroxidevalue of 100. The higher AOM hours of IMC 130 reflects its greater oilstability. Each oil sample was prepared in duplicate. TABLE 11 AOM Hoursof Commercially Processed Canola Oils Process IMC 144 IMC 01 IMC 129 IMC130 Method (Generic) (Low ALA)* (High Oleic) (Example 5) Commercial15-18 30 30 37.5

Example 6

[0087] Another doubled haploid line was identified in the DH₂ generationof the IMC129 X IMC01 cross of Example 0.1 and termed A13.30137.: Theline was self-pollinated and selected as described in Example 1 for fivegenerations. A13.30137 was tested for fatty acid stability and yieldpotential in research plots and strip trials in Idaho, Montana, andWashington. It appeared homogeneous and morphological variation was notobserved during the production of foundation seed.

[0088] A13.30137 matured about two days earlier than IMC130 and wasabout 10 cm shorter than IMC130. The fatty acid composition of A13.30137seeds was determined over 6 generations as described in Example 1 and isshown in Table 12. The yield performance of A12.30137 in research plotsand strip trials is shown in Table 13 for the DH₅ generation and inTable 14 for the DH₇ generation. TABLE 12 Fatty Acid Composition ofA13.30137 from DH₂ to DH₇ Generation Fatty Acid Composition GenerationC_(16:0) C_(18:0) C_(18:1) C_(18:2) C_(18:3) C_(22:1) DH2 3.8 2.1 76.211.5 3.2 0.00 DH3 3.4 1.9 76.8 10.7 3.7 0.00 DH4 3.2 1.8 77.1 9.8 3.50.00 DH5* 3.5 2.2 75.0 10.1 5.3 0.00 DH6* 3.5 1.7 75.3 10.2 4.9 0.00 DH73.6 2.4 76.7 10.2 3.8 0.00

[0089] TABLE 13 Yield Performance of A13.30137 DH₅ Generation Line NameYield (lb/acre) % of Westar Hyola 401 2,171 118 A13.30137 1,879 102IMC129 1,865 102 Westar 1,835 100 Legend 1,764 96 IMC130 1,573 86 Global1,526 83

[0090] TABLE 14 Yield Performance of A13.30137 DH₇ Generation Line NameYield (lb/acre) % of Checks* Cyclone 1,960 131 Hyola 401 1,876 126A13.30137 1,871 122 Delta 1,859 121 Legend 1,804 118 IMC129 1,783 110IMC130 1,625 96 Excel 1,578 93

What is claimed is:
 1. An oil comprising a non-hydrogenated canola oilhaving an oxidative stability of from about 37 to about 40 AOM hours inthe absence of antioxidants.
 2. The oil of claim 1 wherein thepercentage increase of total polar material is about 6.7% after 32 hoursof frying, and about 17.3% after 64 hours of frying.
 3. The oil of claim1 wherein the percentage increase of free fatty acids is about 0.28%after 32 hours of frying and about 0.73% after 64 hours of frying. 4.The oil of claim 1 wherein Lovibond color increase is about 2.4 redafter 32 hours of frying and about 6.4 red after 64 hours of frying. 5.The oil of claim 1 wherein increase in para-anisidine value is about 112absorbance/g after 32 hours of frying and about 125 absorbance/g after64 hours of frying.
 6. The oil of claim 1 wherein the peroxide valueincreases by a maximum of about 24.5 Meq/Kg after accelerated aging for15 days.
 7. The oil of claim 1 wherein the para-anisidine valueincreases by a maximum of about 6.9 absorbance/g after accelerated agingfor 15 days.
 8. A seed comprising a Brassica napus canola varietycontaining an oil of claim
 1. 9. A progeny of a seed of claim
 8. 10. Aseed of claim 8 deposited with the American Type culture Collection andbearing accession number
 75446. 11. A Brassica napus plant linecomprising a canola variety which produces seed oil of claim
 1. 12. Aplant of the line of claim
 11. 13. Progeny of the seed of claim
 10. 14.A seed of claim 8 designated as A13.30137.
 15. Progeny of the seed ofclaim
 14. 16. A Brassica napus line effective for producing seedcontaining an oil of claim 1, said line descending from a cross of lineIMC129 and line IMC01.